Purification and Characterization of a Glutathione Reductase from Phaeodactylum tricornutum

Arias, Diego Gustavo; Márquez, Vanina Elizabet; Beccaria, Alejandro José; Guerrero, Sandra; Iglesias, Alberto A.

doi:10.1016/j.protis.2009.06.001

Cited by 22 publications

(12 citation statements)

References 41 publications

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“…We purified successfully three enzymes from rat heart using the affinity chromatography method. In some reports, the purification of these enzymes has been severally performed using multistep chromatographic methods including affinity, ion‐exchange, and gel filtration chromatography . 2′,5′‐ADP sepharose‐4B affinity chromatography is a special method for purification of NADP + /NADPH‐dependent enzymes such as 6PGD, GR, and G6PD.…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of Glucose 6‐Phosphate Dehydrogenase, 6‐Phosphogluconate Dehydrogenase, and Glutathione Reductase from Rat Heart and Inhibition Effects of Furosemide, Digoxin, and Dopamine on the Enzymes Activities

Adem

Çiftçi

2016

J Biochem & Molecular Tox

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The present study was aimed to investigate characterization and purification of glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase from rat heart and the inhibitory effect of three drugs. The purification of the enzymes was performed using 2',5'-ADP sepharose 4B affinity material. The subunit and the natural molecular weights were analyzed by SDS-PAGE and gel filtration. Biochemical characteristics such as the optimum temperature, pH, stable pH, and salt concentration were examined for each enzyme. Types of product inhibition and Ki values with Km and Vmax values of the substrates and coenzymes were determined. According to the obtained Ki and IC50 values, furosemide, digoxin, and dopamine showed inhibitory effect on the enzyme activities at low millimolar concentrations in vitro conditions. Dopamine inhibited the activity of these enzymes as competitive, whereas furosemide and digoxin inhibited the activity of the enzyme as noncompetitive.

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Section: Discussionmentioning

confidence: 99%

Purification and Characterization of Glucose 6‐Phosphate Dehydrogenase, 6‐Phosphogluconate Dehydrogenase, and Glutathione Reductase from Rat Heart and Inhibition Effects of Furosemide, Digoxin, and Dopamine on the Enzymes Activities

Adem

Çiftçi

2016

J Biochem & Molecular Tox

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“…The pathways of CySS metabolism in plant cells are still unclear; Olm et al [31] suggested that molecules such as GSH, and enzymes, such as glutathione reductase (GR), thioredoxin reductase and glutaredoxins (GRXs), could be responsible for CySS reduction. In protozoa, thioredoxin reductase has such a function [32], but several studies of glutathione reductase (GR) showed that this enzyme is not capable of reducing CySS [33]. There are large gaps in our knowledge of Cys and CySS metabolism in plants, and closing them will further enhance our understanding of plant stress response.…”

Section: Sulphur-containing Amino Acidsmentioning

confidence: 99%

A Central Role for Thiols in Plant Tolerance to Abiotic Stress

Zagorchev

Seal

Kranner

et al. 2013

IJMS

356

154

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Abiotic stress poses major problems to agriculture and increasing efforts are being made to understand plant stress response and tolerance mechanisms and to develop new tools that underpin successful agriculture. However, the molecular mechanisms of plant stress tolerance are not fully understood, and the data available is incomplete and sometimes contradictory. Here, we review the significance of protein and non-protein thiol compounds in relation to plant tolerance of abiotic stress. First, the roles of the amino acids cysteine and methionine, are discussed, followed by an extensive discussion of the low-molecular-weight tripeptide, thiol glutathione, which plays a central part in plant stress response and oxidative signalling and of glutathione-related enzymes, including those involved in the biosynthesis of non-protein thiol compounds. Special attention is given to the glutathione redox state, to phytochelatins and to the role of glutathione in the regulation of the cell cycle. The protein thiol section focuses on glutaredoxins and thioredoxins, proteins with oxidoreductase activity, which are involved in protein glutathionylation. The review concludes with a brief overview of and future perspectives for the involvement of plant thiols in abiotic stress tolerance.

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“…rPsGR activity decreased when the pH value was higher than 8.0. The activity of GR from Phaeodactylum tricornutum was found to decrease significantly above a pH of 8.5 [27]. Moreover, a significant decrease in E. coli S33 GR activity was noticed at pH values below 7.5 [29], and the E. coli GR showed almost complete inactivation at pH values below 5.5 and over 7.5 [28].…”

Section: Biochemical Characteristics Of Rpsgrmentioning

confidence: 93%

“…exhibited optimum activity at 60 • C [18]. The optimal temperature of GR from Phaeodactylum tricornutum [27] and Pennisetum glaucum [12] were 32 and 40 • C, respectively. It is worth noting that the rPsGR activity was 42.2% of its highest activity at 0 • C. However, for GR from Chlamydomonas sp.…”

Section: Biochemical Characteristics Of Rpsgrmentioning

confidence: 99%

A New Cold-Adapted and Salt-Tolerant Glutathione Reductase from Antarctic Psychrophilic Bacterium Psychrobacter sp. and Its Resistance to Oxidation

Wang

Hou

2020

IJMS

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A new glutathione reductase gene (psgr) coding for glutathione reductase (GR) from an Antarctic bacterium was cloned and overexpressed into Escherichia coli (E. coli). A sequence analysis revealed that PsGR is a protein consisting of 451 amino acids, and homology modeling demonstrated that PsGR has fewer hydrogen bonds and salt bridges, which might lead to improved conformational flexibility at low temperatures. PsGR possesses the flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide phosphate (NADPH) binding motifs. Recombinant PsGR (rPsGR) was purified using Ni-NTA affinity chromatography and was found to have a molecular mass of approximately 53.5 kDa. rPsGR was found to be optimally active at 25 °C and a pH of 7.5. It was found to be a cold-adapted enzyme, with approximately 42% of its optimal activity remaining at 0 °C. Moreover, rPsGR was most active in 1.0 M NaCl and 62.5% of its full activity remained in 3.0 M NaCl, demonstrating its high salt tolerance. Furthermore, rPsGR was found to have a higher substrate affinity for NADPH than for GSSG (oxidized glutathione). rPsGR provided protection against peroxide (H2O2)-induced oxidative stress in recombinant cells, and displayed potential application as an antioxidant protein. The results of the present study provide a sound basis for the study of the structural characteristics and catalytic characterization of cold-adapted GR.

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Purification and Characterization of a Glutathione Reductase from Phaeodactylum tricornutum

Cited by 22 publications

References 41 publications

Purification and Characterization of Glucose 6‐Phosphate Dehydrogenase, 6‐Phosphogluconate Dehydrogenase, and Glutathione Reductase from Rat Heart and Inhibition Effects of Furosemide, Digoxin, and Dopamine on the Enzymes Activities

Purification and Characterization of Glucose 6‐Phosphate Dehydrogenase, 6‐Phosphogluconate Dehydrogenase, and Glutathione Reductase from Rat Heart and Inhibition Effects of Furosemide, Digoxin, and Dopamine on the Enzymes Activities

A Central Role for Thiols in Plant Tolerance to Abiotic Stress

A New Cold-Adapted and Salt-Tolerant Glutathione Reductase from Antarctic Psychrophilic Bacterium Psychrobacter sp. and Its Resistance to Oxidation

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